WO2017022887A1 - Système végétal rapporteur de détection de composés aromatiques non dégradables nocifs et leur utilisation - Google Patents

Système végétal rapporteur de détection de composés aromatiques non dégradables nocifs et leur utilisation Download PDF

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WO2017022887A1
WO2017022887A1 PCT/KR2015/011937 KR2015011937W WO2017022887A1 WO 2017022887 A1 WO2017022887 A1 WO 2017022887A1 KR 2015011937 W KR2015011937 W KR 2015011937W WO 2017022887 A1 WO2017022887 A1 WO 2017022887A1
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gene
plant
promoter
growth hormone
pseudomonas putida
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Korean (ko)
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류충민
이혜란
권오석
이승구
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한국생명공학연구원
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing

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  • the present invention relates to a reporter plant system for detecting hardly decomposable hazardous aromatic compounds and uses thereof.
  • non-degradable aromatic compound When a non-degradable aromatic compound is released into nature through a food chain or other pathway, it is usually carcinogenic or acts as an environmental hormone by reacting with a polymer such as DNA or protein in cells with its structural similarity and high reactivity. Examples include dioxins, which are emitted during the combustion of automobile emissions and garbage, which not only cause cancer, but also cause reproductive and immune system abnormalities. Also, DDT (dichloro-diphenyl-trichloroethane) is produced as a pesticide. It acts as an environmental hormone that is harmful to the reproductive system. Hardly degradable aromatic compounds entering nature are classified as environmentally toxic substances to be removed first in developed countries.
  • BTEX compounds refer to benzene, toluene, ethylbenzene and xylene. Recently, due to the rapid growth of industrial activities, a considerable amount of harmful pollutants are released into the natural environment, among which VOCs (volatile BTEX compounds, a representative oil compound of organic compounds, are becoming more contaminated due to leakages and accidents in oil storage and widespread use in industry. Pollution caused by these is widely found in the public drinking water system such as surface water source, ground water source, and treated drinking water. Most VOCs are found to be toxic organic chemicals that cause carcinogenicity. Directives and legislation are on the rise.
  • chromatography-mass spectrometry In order to measure such BTEX, chromatography-mass spectrometry (GC-MS) and the like have been conventionally used as an instrument chemical method, but such a method is expensive, takes a long time by complicated processing, and has no expertise. Silver is not easy to use and also requires heavy equipment, there was a limitation in use that can not be used in the field (Korean Patent Publication No. 10-2004-0076292).
  • BTEX compounds benzene, toluene, ethylbenzene, xylene
  • TNT 2,4,6-trinitrotoluene
  • DNT 2,4-Dinitrotoluene
  • Korean Patent No. 1430685 discloses an artificial biosensor for detecting a degradable harmful aromatic compound and a manufacturing method thereof
  • Korean Patent No. 0539142 discloses a biosensor for detecting a BT extract and a method of manufacturing the same. have.
  • the reporter plant system for detecting the hardly degradable harmful aromatic compound of the present invention and its use.
  • the TodST-IAA module operates in the presence of toluene to transform plant to produce plant growth hormone, and includes a DR5 promoter and a GUS gene which recognizes the same.
  • RNAi expression vector comprising an expression vector or a DR5 promoter and a ChlH (Mg-chelatase H subunit) gene
  • ChlH Mg-chelatase H subunit
  • the present invention is 1) Pseudomonas putida derived from todS gene, Pseudomonas putida derived from todT gene, Pseudomonas putida derived from the promoter (promoter) and plant growth hormone (plant growth hormone) Transformed rhizobacteria for the production of plant growth hormone in the presence of a hardly degradable noxious aromatic compound characterized by being transformed with an expression vector comprising a gene: and 2) a promoter and reporter gene specifically recognized by the plant growth hormone
  • a reporter plant system for detecting hardly decomposable harmful aromatic compounds in soil comprising a plant transformed with an RNAi expression vector comprising a promoter and a pigment generating gene that the expression vector or plant growth hormone specifically recognizes.
  • the present invention provides a method for producing a reporter plant system for detecting hardly decomposable harmful aromatic compounds in the soil.
  • the present invention also provides a method for detecting a hardly decomposable harmful aromatic compound in soil using a reporter plant system for detecting a hardly decomposable harmful aromatic compound in soil.
  • the present invention can provide a reporter plant system which is very useful for detecting hardly decomposable harmful aromatic compounds including toluene, and can more sensitively and accurately detect the hardly decomposable hazardous aromatic compounds than in the prior art.
  • it is possible to recognize the direct simplicity rather than the indirect external recognition system of the area where people are not easily accessible, and it can be useful for developing a system that can recognize toxic substances in real life without using chemical / physical equipment. have.
  • Figure 1 shows a schematic diagram of toluene reporter plant production using Pseudomonas putida TodST-IAA module and DR5-GUS system of the present invention.
  • FIG. 2 shows the biosynthetic pathway of plant growth hormone IAA (Indole-3-acetic acid) produced by bacteria.
  • IAA Indole-3-acetic acid
  • the purple arrow is the IAM (indole-3-acetamide) pathway and the red arrow is the IPyA (Indole-3-pyruvate) pathway.
  • Figure 3 shows the IAA production capacity of Pseudomonas putida 06iaaMH, 30iaaMH and ipdC strain of the present invention when treated with toluene.
  • Figure 4 shows the results of GUS expression of Arabidopsis plants using IAA production capacity of Pseudomonas putida 06iaaMH, 30iaaMH and ipdC strain of the present invention with or without toluene.
  • Figure 5 shows the root colonization capacity (root colonization capacity) of the Pseudomonas putida strain of the present invention.
  • Figure 6 shows a schematic diagram of toluene reporter transgenic plant production using Pseudomonas putida TodST-IAA module and Ch1H-RNAi system of the present invention.
  • FIG. 7 shows a schematic diagram of a method for producing a pK7_DR5_Ch1H vector of the present invention.
  • Figure 8 shows the RNAi phenotype of the Ch1H gene when treated with Agrobacterium tumefasiens strains and IAA standards transformed with RNAi expression vector on the leaves of tobacco plants Nicotiana benthamiana with or without toluene. It is shown. Mock is a negative control treated with nothing.
  • RNAi expression vectors on a leaf of the plant conversion was Agrobacterium Tome Pacific Enschede (Agrobacterium tumefasiens) strain, IAA-producing strain of Pseudomonas footage is (Pseudomonas putida) O6iaaMH strain and Pseudomonas When treated with Pseudomonas chlororaphis strain and IAA standards, the RNAi phenotype of the Ch1H gene is shown. Water control is a negative control with no treatment.
  • RNAi expression vectors on a leaf of the plant conversion was Agrobacterium Tome Pacific Enschede (Agrobacterium tumefasiens) strain, IAA-producing strain of Pseudomonas footage is (Pseudomonas putida) O6iaaMH strain and Pseudomonas Chlorophyll content is shown when treated with Pseudomonas chlororaphis strain and IAA standards. Water control is a negative control with no treatment.
  • the present invention is a.
  • a plant transformed with an expression vector comprising a promoter and a reporter gene specifically recognized by the plant growth hormone or an RNAi expression vector comprising a promoter and a pigment generating gene specifically recognized by the plant growth hormone Provided is a reporter plant system for detecting hardly decomposable harmful aromatic compounds in soil.
  • the todS gene preferably has a nucleotide sequence of SEQ ID NO: 7, but is not limited thereto.
  • the todT gene preferably has a nucleotide sequence of SEQ ID NO: 8, but is not limited thereto.
  • the promoter of the todX gene preferably has a nucleotide sequence of SEQ ID NO: 9, but is not limited thereto.
  • the promoters of the todS gene, the todT gene, and the todX gene are composed of nucleotide sequences in which one or several bases are added, deleted, or substituted as long as they have the same activity in the nucleotide sequences of SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, respectively. It is preferred, but not limited to.
  • the promoters of the todS gene, the todT gene, and the todX gene are at least 80% homology, more specifically at least 90% homology, most specifically 95%, 96% to the nucleotide sequences of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, respectively. It is preferably composed of a base sequence having a homology of at least 97%, 98%, 99% or 99.5%, but is not limited thereto.
  • the plant growth hormone is preferably IAA (indole acetic acid), cytokinin (cytokinin), gibberellin (gibberellin), ethylene (ethylene), abscisic acid (brassinosteroid), etc., but is not limited thereto. Do not.
  • the promoter specifically recognized by the plant growth hormone is preferably a DR5 promoter, a cytokinin receptor promoter, an ethylene receptor promoter, or a pathogenesis-related 1 promoter, but is not limited thereto. .
  • the reporter gene is composed of green fluorescent protein (GFP), alkaline phosphatase, luciferase, luciferase, beta-glucuronidase (GUS) and beta-galactosidase genes. It is preferably any one selected from the group, more preferably beta-glucuronidase (GUS) gene, but is not limited thereto.
  • GFP green fluorescent protein
  • GUS beta-glucuronidase
  • the pigment generating gene is preferably a ChlH (Mg-chelatase H subunit) gene or a PDS (phytoene desaturase) gene, but is not limited thereto.
  • the rhizosphere bacteria are Pseudomonas putida , Paenibacillus polymyxa E681, Bacillus subtilis GB03, Bacillus pumilus INR7 is preferred, but not limited thereto.
  • the hardly decomposable hazardous aromatic compound may be a BTEX compound (benzene, toluene, ethylbenzene, xylene) or toluene compound (for example, 2,4,6-trinitrotoluene (2). , 4,6-trinitrotoluene, TNT)), more preferably toluene, but is not limited thereto.
  • BTEX compound benzene, toluene, ethylbenzene, xylene
  • toluene compound for example, 2,4,6-trinitrotoluene (2). , 4,6-trinitrotoluene, TNT
  • vector is used to refer to a DNA fragment (s), a nucleic acid molecule, that is delivered into a cell. Vectors can replicate DNA and be reproduced independently in host cells.
  • carrier is often used interchangeably with “vector”.
  • expression vector refers to a recombinant DNA molecule comprising a coding sequence of interest and a suitable nucleic acid sequence necessary to express a coding sequence operably linked in a particular host organism. Promoters, enhancers, termination signals and polyadenylation signals available in eukaryotic cells are known.
  • Ti-plasmid vectors which, when present in a suitable host such as Agrobacterium tumerfaciens, can transfer part of themselves, the so-called T-region, into plant cells.
  • a suitable host such as Agrobacterium tumerfaciens
  • Another type of Ti-plasmid vector (see EP 0 116 718 B1) is used to transfer hybrid DNA sequences to protoplasts from which current plant cells or new plants can be produced that properly insert hybrid DNA into the plant's genome. have.
  • a particularly preferred form of the Ti-plasmid vector is the so-called binary vector as claimed in EP 0 120 516 B1 and US Pat. No. 4,940,838.
  • viral vectors such as those which can be derived from double stranded plant viruses (eg CaMV) and single stranded viruses, gemini viruses, etc.
  • CaMV double stranded plant viruses
  • gemini viruses single stranded viruses
  • it may be selected from an incomplete plant viral vector.
  • the use of such vectors can be advantageous especially when it is difficult to properly transform a plant host.
  • the expression vector will preferably comprise one or more selectable markers.
  • the marker is typically a nucleic acid sequence having properties that can be selected by chemical methods, and all genes that can distinguish transformed cells from non-transformed cells. Examples include herbicide resistance genes such as glyphosate, phosphinothricin and glufosinate, kanamycin, G418, bleomycin, hygromycin, There are antibiotic resistance genes such as chloramphenicol, but are not limited thereto.
  • the promoter may be, but is not limited to, CaMV 35S, actin, ubiquitin, pEMU, MAS or histone promoter.
  • promoter refers to a region of DNA upstream from a structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription.
  • a "plant promoter” is a promoter capable of initiating transcription in plant cells.
  • a “constitutive promoter” is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Constitutive promoters may be preferred in the present invention because selection of the transformants may be made by various tissues at various stages. Thus, the constitutive promoter does not limit the selection possibilities.
  • terminators can be used, for example nopalin synthase (NOS), rice ⁇ -amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens (Agrobacterium) terminators of the octopine gene of tumefaciens, but are not limited thereto.
  • NOS nopalin synthase
  • rice ⁇ -amylase RAmy1 A terminator Phaseoline terminator
  • Agrobacterium tumefaciens (Agrobacterium) terminators of the octopine gene of tumefaciens but are not limited thereto.
  • terminators With regard to the need for terminators, such regions are generally known to increase the certainty and efficiency of transcription in plant cells. Therefore, the use of terminators is highly desirable in the context of the present invention.
  • Plant transformation refers to any method of transferring DNA to a plant. Such transformation methods do not necessarily have a period of regeneration and / or tissue culture. Transformation of plant species is now common for plant species, including both dicotyledonous plants as well as monocotyledonous plants. In principle, any transformation method can be used to introduce hybrid DNA according to the invention into suitable progenitor cells. The method is based on the calcium / polyethylene glycol method for protoplasts (Krens et al., 1982, Nature 296: 72-74; Negrutiu et al., 1987, Plant Mol. Biol. 8: 363-373), electroporation of protoplasts ( Shillito et al., 1985, Bio / Technol.
  • step 2) transforming the expression vector of step 1) to the rhizosphere bacteria to prepare a transformed rhizosphere bacterium for plant growth hormone production in the presence of a hardly decomposable harmful aromatic compound;
  • RNAi expression comprising the expression vector comprising a promoter and a reporter gene that the plant growth hormone specifically recognizes the transformed rhizosphere bacteria of step 2) or a promoter and a pigment generating gene that the plant growth hormone specifically recognizes
  • a method for producing a reporter plant system for detecting a non-degradable harmful aromatic compound in soil comprising the step of treating the plant transformed with the vector.
  • each gene, plant growth hormone, rhizosphere bacteria, reporter gene, pigment generating gene and the like are as described above.
  • RNAi expression vector comprising an expression vector comprising a promoter and a reporter gene specifically recognized by the plant growth hormone or a promoter and a pigment generating gene specifically recognized by the plant growth hormone in the test sample. Planting step;
  • the color change of the transgenic plant can be confirmed in comparison with the non-transformed plant, but is not limited thereto. That is, when toluene is present in the test sample, the plant transformed with the expression vector including the reporter gene may be colored through expression of the reporter gene, compared to the non-transformed plant, and RNAi including the pigment generating gene. Plants transformed with the expression vector may have a problem in the production of chloroplast pigment, and the plant may turn yellow. Therefore, the presence or absence of toluene can be detected through the color change of the transgenic plant.
  • Pseudomonas putida ( Pseudomonas putida ) Fabrication of TodST-IAA Module
  • the TodST module using the two-component system of microorganisms was used.
  • IAA Indole-3-acetic acid
  • TodT-BOX the artificial biosensor for detecting degradable harmful aromatic compound and its manufacturing method
  • Pseudomonas putida KT2440 strain was used (FIG. 1).
  • Auxin (auxin) is a representative plant growth hormone affects the growth of the cells, some bacteria in the plant root area to produce and secrete auxin, which can be absorbed by the plant and used for growth.
  • IAA Indole-3-acetic acid
  • IAM indole-3-acetamide pathway
  • IPyA Indole-3-pyruvate pathway
  • IAAld indole-3-acetaldehyde
  • IPDC Indolepyruvate decarboxylase
  • iaaM , iaaH and ipdC genes In order to produce a Pseudomonas (Pseudomonas) in and azo RY rilrum (Azospirillum) in strains with the expression vectors to produce high levels of IAA, iaaH and iaaM gene from Pseudomonas chloro lapis (Pseudomonas chlororaphis) O6 and 30-84 strains The ipdC gene was obtained from an Azospirillum brasilense Cd strain to prepare an expression vector.
  • Pseudomonas chloro lapis (Pseudomonas chlororaphis) O6 and 30-84 strains, and azo RY rilrum bra chamber lances (Azospirillum brasilense) inoculated with the respective strain Cd KB (King's B) 3ml broth and incubated at 30 °C 24 hours. Then, the cells were collected by centrifugation and genomic DNA was extracted using NanoHelix PureHelix Genomic DNA Prep Kit (Column Type). Since genomic DNA as a template amplified iaaM , iaaH and ipdC using Roche's Expand High Fidelity PCR system.
  • Pseudomonas putida KT2440 strain (pBBRBB_TodST) was transformed using Bio-Rad's Gene Pulser Xcell Electroporation System, and colonies growing on LB agar medium containing 50 ⁇ g / ml of gentamicin were selected and Pseudomonas putida. ) O6iaaMH, Pseudomonas putida 30iaaMH and Pseudomonas putida ipdC strains were obtained.
  • the absorbance was then measured at 530 nm using an UV spectrophotometer, and a standard curve was prepared using IAA (indole-3-acetic acid, Sigma, USA) as a standard and compared with the IAA standard curve. OD value was converted into IAA concentration.
  • the footage is Pseudomonas (Pseudomonas putida) ip dC, O6iaa MH and MH iaa 30 strain having a TodST-IAA module as set forth in 3, when processing the toluene, about 31 ⁇ g / ml respectively, 38 ⁇ g / ml And 20 ⁇ g / ml of IAA, but did not produce IAA when not treated with toluene.
  • the Pseudomonas chlororaphis O6 strain producing IAA it was confirmed that the color reaction occurred regardless of toluene treatment.
  • the cell suspension was then irrigated 10 ml each to Arabidopsis thaliana DR5-GUS , followed by 100 ⁇ M of toluene after 3 days. After 3 days of treatment, Arabidopsis thaliana DR5-GUS was extracted and stained with GUS to confirm GUS expression. Plants were placed in GUS staining reagent (100mM NaH 2 PO 4 , 5mM Potassium Ferricyanide, 5mM Potassium Ferrocyanide, 10mM EDTA, 0.1% Triton X-100, 5mg / ml X-Gluc) and soaked for 12 hours at 37 ° C. Were removed and washed until the plant became white in 50% ethanol.
  • GUS staining reagent 100mM NaH 2 PO 4 , 5mM Potassium Ferricyanide, 5mM Potassium Ferrocyanide, 10mM EDTA, 0.1% Triton X-100, 5mg / ml
  • Pseudomonas putida was produced to observe the root zone fixation to confirm whether the plant exists while maintaining the interaction in the plant roots.
  • a wild tobacco species Nicotiana benthamiana was selected as a model plant, and the target gene was selected from tobacco Chl H (magnesium protoporphyrin chelatase H subunit).
  • Chl H is an important enzyme that chelates magnesium in the chloroplasts of plants. When RNAi of this gene occurs, it causes problems in the production of pigments of chloroplasts and turns the plants yellow.
  • Chl H gene fragment was obtained using the primer set of Chl H in Table 1 below. Since each primer contains a position capable of recombination with a vector, a construct was constructed to express Chl H by recombinantly in an RNAi-enabled plasmid in Nicotiana benthamiana .
  • Chl H RNAi vectors were prepared for pK7GWIWG2-I, a binary vector used for hairpin RNA expression.
  • the bacteria's two-component system recognizes external VOCs (Volatile organic compounds) in TodS, which causes phosphorylation to activate TodT, moves into the nucleus, and binds to the TodT box.
  • DR5 promoter reported as an auxin response element
  • RNAi vectors Genomic DNA was extracted from transgenic Arabidopsis seedlings inserted with DR5 :: GUS, and the gene fragment of about 300bp corresponding to the DR5 sequence and the -46 CaMV minimal promoter region was extracted using the primers of Table 1 below. Amplified.
  • the attR-ChlH-attR-intron portion was amplified with the primers of Table 1 below, and amplified so as to be connected to the DR5 promoter, thereby obtaining an insert of about 1.2 kb. . Since the amplification was carried out with a primer including a position capable of recombination with the vector, pK7_DR5_ccdB vector was constructed through recombination.
  • PK7GWIWG2 (I) containing the ChlH gene was obtained through the gateway cloning system BP reaction to obtain the pDONR-ChlH vector, which is an RNAi vector containing the DR5 promoter and Chl H through the LR reaction with the pK7_DR5_ccdB vector.
  • pK7_DR5_ChlH was obtained (FIG. 7).
  • the constructed pK7_DR5_Ch1H vector was transformed into the Agrobacterium tumefaciens GV2260 strain, and then Agrobacterium tumefaciens ( A. tumefaciens ) cell suspension on the leaves of Nicotiana benthamiana . Direct infiltration was performed. Then, after 1-2 days, IAA standards were infiltrated at various concentrations of 100 nM, 10 ⁇ M, 100 ⁇ M and 1 mM. As a result, as shown in FIG. 8, the Chl H phenotype was observed to change the color of the leaf only at the intersection of the IAA standard treatment group and the Agrobacterium tumefaciens treatment group. Part did not show the Chl H phenotype.
  • Chl H RNAi turned yellow regardless of all treatments, but in DR5- chl H, the Pseudomonas putida O6iaaMH treatment, Pseudomonas putida treated with toluene, Pseudomonas chlorolapi treatment and IAA treatment only It was observed that the cross section treated with the bacterium tumefaciens strain turned yellow.
  • Chl H RNAi showed low chlorophyll content regardless of all treatments, but DR5- chl H showed high chlorophyll content in toluene-treated treatments, but Pseudomonas treated with toluene. Chlorophyll content was low only in Pseudomonas putida and IAA treatments.
  • IAA generated by Pseudomonas putida strain with TodST-IAA module in the presence of toluene was able to confirm the operation of the phyto-sensor system in which chl H RNAi occurred and changed the phenotype.

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Abstract

La présente invention concerne un système végétal rapporteur de détection de composés aromatiques non dégradables nocifs et une utilisation de celui-ci. La présente invention peut fournir un système végétal rapporteur qui est très utile pour la détection de composés aromatiques non dégradables nocifs, comprenant le toluène, et qui est capable de détecter les composés aromatiques non dégradables nocifs de manière plus sensible et plus précise que l'art antérieur. La présente invention rend également possible la reconnaissance directe et simple d'une zone inaccessible aux êtres humains plutôt qu'à l'aide d'un système de reconnaissance externe indirect, et peut être efficacement utilisée pour élaborer un système capable de reconnaître les substances toxiques dans la vie réelle sans utiliser d'appareil chimique ou physique.
PCT/KR2015/011937 2015-08-06 2015-11-06 Système végétal rapporteur de détection de composés aromatiques non dégradables nocifs et leur utilisation WO2017022887A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107365729A (zh) * 2017-08-25 2017-11-21 山东烟草研究院有限公司 一种多粘类芽孢杆菌及其应用
CN113373102A (zh) * 2021-07-05 2021-09-10 青岛农业大学 一种检测爆炸物的新型光合细菌及其制备方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20010032491A (ko) * 1997-11-25 2001-04-25 유티-배텔, 엘엘씨 생체 발광 생체 리포터 집적회로
WO2003100084A1 (fr) * 2002-05-23 2003-12-04 National Research Council Of Canada Nouveau biodetecteur
CN101619341A (zh) * 2008-06-30 2010-01-06 财团法人生物技术开发中心 用于有机污染物浓度与降解效能检测的转化株及其检测方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101430685B1 (ko) * 2011-11-17 2014-08-14 한국생명공학연구원 난분해성 유해 방향족 화합물 감지용 인공바이오센서 및 이의 제조방법

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20010032491A (ko) * 1997-11-25 2001-04-25 유티-배텔, 엘엘씨 생체 발광 생체 리포터 집적회로
WO2003100084A1 (fr) * 2002-05-23 2003-12-04 National Research Council Of Canada Nouveau biodetecteur
CN101619341A (zh) * 2008-06-30 2010-01-06 财团法人生物技术开发中心 用于有机污染物浓度与降解效能检测的转化株及其检测方法

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BUSCH, ANDREAS ET AL.: "Catabolite Repression of the TodS/TodT Two-Component System and Effector-Dependent Transphosphorylation of TodT as the Basis for Toluene Dioxygenase Catabolic Pathway Control", JOURNAL OF BACTERIOLOGY., vol. 192, no. 16, 2010, pages 4246 - 4250, XP055362792 *
LEE, HAE - RAN ET AL.: "The Engineered Rhizosphere Bacterium Help Plant Sense a Hazards Chemical", 10TH INTERNATIONAL PGPR WORKSHOP, 16 June 2015 (2015-06-16), pages 46 - 47 *
LEE, HAE - RAN ET AL.: "The Engineered Rhizosphere Bacterium Help Plant Sense a Hazards Chemical", RHIZOSPHERE4, 21 June 2015 (2015-06-21), pages 317 - 318 *
LEE, SEUNG GU ET AL.: "Next generation BioGreen 21 Business-Designing Bio-parts and Circuits for Sentinel Plant Systems", REPORT OF SYSTEMS & SYNTHETIC AGROBIOTECH CENTER, 7 November 2014 (2014-11-07), pages 1 - 37 *
LEE, SEUNG GU ET AL.: "Phyto-Sensor Development Based on Synthetic Biology", RURAL DEVELOPMENT ADMINISTRATION, STUDY REPORT OF NEXT GENERATION BIOGREEN 21 BUSINESS, 23 February 2013 (2013-02-23), pages 1 - 89 *

Cited By (4)

* Cited by examiner, † Cited by third party
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CN107365729A (zh) * 2017-08-25 2017-11-21 山东烟草研究院有限公司 一种多粘类芽孢杆菌及其应用
CN107365729B (zh) * 2017-08-25 2020-10-16 山东烟草研究院有限公司 一种多粘类芽孢杆菌及其应用
CN113373102A (zh) * 2021-07-05 2021-09-10 青岛农业大学 一种检测爆炸物的新型光合细菌及其制备方法和应用
CN113373102B (zh) * 2021-07-05 2022-09-09 青岛农业大学 一种检测爆炸物的光合细菌及其制备方法和应用

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